Anti ocn

Using a DIW 3D‐Bioplotter System (EnvisionTEC, Germany), a cube structure (10 × 10 × 2.5 mm) with an inner strand distance of 0.9 mm and inner strand angles of 0° and 90° was built to mimic alveolar bone. DFCs were mixed with two bioinks at a density of 5 × 10⁶ cells mL⁻¹. The needle offset along the z‐direction was 210 µm, which was 80% of an ideal strand diameter (260 µm) for each printed layer. The printing pressure was set as 90 kPa for the GelMA bioink and 150 kPa for the GelMA+5dECM bioink. The printing speed was 12 mm s⁻¹, and the printed samples were finally exposed to 405 nm UV light for 30 s.
After culturing for 1, 3, and 7 days in vitro, cell viability was evaluated using a Live/Dead Viability/Cytotoxicity kit (KeyGEN, China). Seven fields were randomly selected from each group for quantitative analysis. To detect osteogenic differentiation in the alveolar bone module, IF staining was performed at 7 and 14 days. The primary antibodies included anti‐ALP (1:100 dilution, Abcam, UK), anti‐Runx2 (1:100 dilution, Abcam, UK) and anti‐OCN (1:200 dilution, Zenbio, China). All samples were examined under a laser scanning confocal microscope (Olympus, Japan).

For histology and immunohistochemistry analysis, sections of 6 µm were made for hematoxylin/eosin staining. Immunohistochemical staining was performed as previously described 35 (link). Primary anti-DSPP (1/200; Zen Bioscience), anti-DMP1 (1/200; Santa Cruz Biotechnology), and anti-OCN (1/200; Zen Bioscience) antibodies were used. Detection of crossreacted secondary antibodies was performed using the DAB kit (Gene Tech, China). Accordingly, quantitative analysis of the staining was conducted by measuring the average optical density using the ImageJ software. At least 3 serial sections from each sample were examined, and at least 3 samples were included in each group.